The invention relates to a method of preventing and treating diseases and conditions of mammals associated with the adhesion, metastatic and coronary cascades comprising applying a composition of complex carbohydrates and essential oils topically, orally or mucosally on a repeated basis. The invention also encompasses a method of preventing and treating diseases and conditions associated with the adhesion, metastatic and coronary cascades comprising orally or mucosally applying complex carbohydrates as the sole active ingredient.
Complex carbohydrates, for purposes of this invention are defined as any polymer comprising more than two sugar moieties including such classes of compounds as polysaccharides and oligosaccharides. Polysaccharides include mucopolysaccharides and mannans whereas oligosaccharides are comprised of branched polysaccharides such as sialylated sugars including milk sugars.
Mucopolysaccharides are glycosaminoglycans which can be obtained from numerous sources (e.g. rooster combs, trachea, umbilical cords, skin, articular fluids and certain bacteria such as Streptococci spp). Most glycosaminoglycans (hyaluronic acid, chondroitin sulfates A, B, and C, heparin sulfate, heparin, keratan sulfate, dermatan sulfate, etc.) are composed of repeating sugars such as n-acetylglucosamine, glucuronic acid and n-acetyl galactosamine (these are known as non-sulfated glycosaminoglycans). If such glycosaminoglycans contain sulfur groups they are known as sulfated glycosaminoglycans.
Mannans are mannose-based polysaccharides which are normally extracted from plants. The most noteworthy is acemannan which is a beta 1,4-linked acetylated mannan extracted from the Aloe Vera plant (Aloe barbadensis Miller). This plant has been thought for centuries to have certain healing powers. Not until the 1980s was the active ingredient isolated and proven to have an effect on the immune system (see J. Pharm. Sci., 73 (1), January, 1984). Sialylated sugars are oligosaccharides which contain sialyl groups (e.g. sialic acid) and often contain fucose. Sialyl Lewisx and its derivatives are examples from this group (Tyrell et al, Proc. Natl. Acad. Sci. USA, 88, November 1991). At present, this oligosaccharide is so difficult to prepare/obtain that the cost is prohibitive and limits research activities to determine its mechanism of action. Some of the milk sugars (also called hexaoses) are also incorporated in this general class of compounds. Examples of these are difucosyllacto-N-hexaose a and b, Disialyl-monofucosyllacto-N-hexaose and monofucosyllacto-N-hexsaose I, II, and II (obtainable from Oxford Glycosystems, Inc.).
Heparin, hyaluronic acid and chondroitin sulfate are the most studied complex carbohydrates. They fall in the class called mucopolysaccharides or glycosaminoglycans. Heparin has been used for a number of years as an anticoagulant. Hyaluronic acid has been used therapeutically since the 1970s as a replacement for the vitreous humor of the eye post surgery and, more recently, as replacement for joint fluid in arthritic joints. An extensive discussion of its various utilities is found in U.S. Pat. No. 4,141,973 to Balazs. The mode of action for hyaluronic acid injected directly into joints for treatment of arthritis has been proposed to be lubrication and replacement of the degraded joint fluid with highly viscous hyaluronic acid (see J. Bone Jt. Surg. 54A, 1972). High molecular weight (>1,000,000 daltons) and high viscosity have been reported to be critical. (For purposes of this application, all molecular weights are expressed as daltons. The unit designation will not be added hereafter.)
In the 1980s, it was discovered that chondroitin sulfate, or polysulfated glycosaminoglycan (known by its commercial name as ADEQUAN□) could be injected intramuscularly for reduction of pain and inflammation associated with arthrosis of horses. The mechanism of action of this glycosaminoglycan has been speculated to be inhibition of certain degradative enzymes present in the joint fluid which are up-regulated by trauma.
In the 1990s, chondroitin sulfate had developed into a popular nutritional supplement being used extensively to treat joint disease. Such treatment requires oral doses between 1000 and 3000 mg/day of for humans. Even with these high doses, relief from joint pain often takes 6-9 months.
In 1989, it was discovered that intravenous, intramuscular or subcutaneous delivery of hyaluronic acid could reduce the pain of arthritis (U.S. Pat. No. 4,808,576 by Schultz et al) when the hyaluronic acid was delivered remote to the site of the arthritis (not into the joint). This patent specifically states that the hyaluronic acid is administered remote to the site and that the hyaluronic acid must be of high purity (>99% pure hyaluronic acid). Schultz et al. does not disclose or suggest the use of hyaluronic acid in combination with essential oils, use of other complex carbohydrate macromolecules, oral application or mucosal application. Schultz et al. specifically teaches away from use of low purity complex carbohydrates. By low purity is meant complex carbohydrates that would be considered food grade or cosmetic grade, which could be <98% pure and could contain such contaminants as endotoxins, lipoteichoic acids, proteins, nucleic acids, etc. The low purity hyaluronic acid or salt thereof useful in the present invention (<98% pure hyaluronic acid) can be of a cosmetic grade or food grade which can contain up to 5% contaminants. Such material would not pass the owl monkey eye test used to select high purity hyaluronic acids and salts thereof (described by Balazs in U.S. Pat. No. 4,141,973) in that it would produce an inflammatory response in the eye. It also would not pass the horse joint injection test described by Schultz et al (U.S. Pat. No. 4,808,576). However, it does not produce a reaction when applied to the skin or mucous membranes of mammals including humans, dogs, cats, horses, cattle, swine, rabbits, guinea pigs and mice.
The importance of high molecular weight for effectiveness of hyaluronic acid in the treatment of arthritis is emphasized by Balazs (U.S. Pat. No. 4,141,973) and in a publication by Howard and McIlraith (see The Compendium, 15(3), March 1993) who summarize several clinical studies conducted to determine the most efficacious molecular weight range of hyaluronic acid injected intra-articularly to treat traumatic arthritis in horses. The conclusion from these studies is that hyaluronic acid with a molecular weight below 1×106 is not as effective as hyaluronic acid with a molecular weight above this value. More recently, della Valle et al (U.S. Pat. No. 5,166,331) claimed that there are two distinct pharmacologically active molecular weight ranges of hyaluronic acid or salts thereof. These moieties are utilized separately (purified one from the other) and defined as 50,000-100,000 (Hylastine) and 500,000-730,000 (Hylectin). Hylastine is specified for use in wound healing while Hylectin is specified for use in ocular surgery.
Whereas Balazs (U.S. Pat. No. 4,141,973), Schultz (U.S. Pat. No. 4,808,576) and della Valle (U.S. Pat. No. 5,166,331) all specify use of highly purified hyaluronic acid and whereas Balazs (U.S. Pat. No. 4,141,973) discards the fractions containing hyaluronic acid or their salts having molecular weights less than 750,000; and whereas della Valle (U.S. Pat. No. 5,166,331) discards impurities having molecular weights less than 30,000 and does not use hyaluronic acid with molecular weights between 100,000 and 500,000 and, thus, specifies use of clearly-defined molecular weights of hyaluronic acid for topical or ocular use; and whereas Schultz prefers use of hyaluronic acid with a molecular weight between 1.2×106 and 4.0×106 in topical formulations, we have discovered that all molecular weights of complex carbohydrates such as hyaluronic acids or salts thereof and all purities of these compounds are useful in topical, oral or mucosal preparations for the treatment of numerous diseases and conditions.
The most recent studies on hyaluronic acid discuss treatment of various types of cancer with very large doses of this macromolecule (Falk, WO 97/40841). The Falk application suggests that doses should exceed 750 mg. per 70 kg person, preferably, exceeding 1 g. per 70 kg person. Such doses are given intermittently post diagnosis and are not suggested to be preventative or administered in low doses. Additionally, it is clear that the sodium hyaluronate of Falk needs to be pure enough for injection even though oral administration is used in addition to intravenous injection.
Essential oils are natural components of plants and animals that are extracted by various methods known to the art. They are generally very complex, containing numerous compounds (see Perfumer and Flavorist, 17, November/December 1992). More recently, some of the essential oils have been chemically synthesized. Most uses of these oils are as flavorings for foods and candies and as bath, cosmetic and perfume ingredients to provide pleasant aromas. Several of the essential oils (i.e. Menthol, Eucalyptus Oil, Camphor, Peppermint Oil and Wintergreen Oil) are currently used in over-the-counter topical preparations such as BenGay, Mineral Ice, Flexall 454, etc. at concentrations as high as 50%. These topical medications claim pain relief but, according to FDA, act to relieve pain by producing a counterirritation, not by penetrating the skin and acting systemically to reduce inflammation and swelling which are the causes of pain.
The Adhesion Cascade was first described in the early 1990s. In a summary by Adams and Shaw (The Lancet, 343, Apr. 2, 1994) the adhesion cascade which is stimulated when trauma occurs is divided into four sequential steps of tethering, triggering, strong adhesion and motility. Tethering interactions are mediated by a family of three lectin-like carbohydrate-binding molecules (selectins). These interactions are strong enough to cause the leukocytes to roll along the blood vessel walls to the site of trauma instead of flowing freely through such vessels, but not strong enough to cause these leukocytes to slow down. The triggering response is stimulated by factors such as cytokines and mediated by adhesion molecules called integrins. Integrins, by themselves, do not bind well to epithelium. However, when activated, integrins promote strong adhesion of the leukocyte to the epithelial surface. Leukocytes bind to the epithelial cells via their receptor sites such as CD44, CD31, etc. During strong adhesion, the interaction of these integrins with their ligands on the surface of the leukocytes are responsible for cessation of movement and flattening of the leukocyte. Finally, a process involving VCAM-1 and LFA-1 and other such integrins allows leukocytes to pass between endothelial cell junctions and into the tissue that has been traumatized. Collection of leukocytes at the site of trauma produces inflammation which is then followed by pain or other sequelae.
The present invention is based upon the premise that complex carbohydrates, including but not limited to glycosaminoglycans, bind to the receptor sites on leukocytes blocking their ability to tether to the blood vessel walls thus inhibiting the motility and interrupting the Adhesion cascade.
The metastatic cascade is very similar to the adhesion cascade. It has been proposed that tumor cells of all types contain CD44 receptor sites on their surface. These CD44 receptor sites appear to be involved in metastasis functioning similar to the receptor sites on leukocytes—tethering the tumor cells to the blood vessel wall and providing the motility necessary for movement from one site to another in the mammalian body. Once again, it is the premise of the present invention that complex carbohydrates, including but not limited to glycosaminoglycans, bind to the receptor sites on tumor cells blocking their ability to tether to the blood vessel walls and inhibiting the motility which, in turn, interrupts the potential for metastasis.
A Coronary cascade has recently been described in the Harvard Health Letter (December 1999, pg. 4-5). This cascade leads to the development of heart disease and stroke by causing plaque formation in the blood vessels. The theory is based on the premise that there are stable and unstable plaques produced on blood vessel walls. Unstable plaques are “swarming with T cells and macrophages” causing inflammation and make these plaques unstable. The T cells are described as sending macrophages a signal to release a protein called tissue factor which “spills out and encounters circulating blood, attracting platelets and triggers formation of a clot that quickly blocks up the artery”. The compositions of the present invention are believed to inhibit the macrophages from infiltrating into the unstable plaques, thus preventing and treating heart disease and stroke.
It is unexpected that complex carbohydrates of the present invention could be administered topically, orally or mucosally in low doses to inhibit the various cascades preventing and treating such a broad spectrum of diseases and conditions.